Induction of tumor necrosis factor α and interleukin-1β in subcutaneously implanted chamber by lipopolysaccharide

Lipopolysaccharide (LPS) is the major component of the outermost membrane of Gram-negative bacteria and is considered to be one of the major virulence factors of these bacteria. While the effect of systemic injection of LPS is well characterized, the characterization of cytokine secretion in response to local injection of LPS is lacking. The present study was designed to determine the local production of tumor necrosis factor α (TNFα) and interleukin-1β (IL-1β) over a 4 day period following injection of LPS into subcutaneous implanted chambers in mice. Mice were challenged by a single or repeated injection of Salmonella typhosa LPS into the chambers. Chamber fluids were aspirated at different time intervals and were used for assessment of leukocyte and cytokine levels. A single injection of LPS was found to induce cell influx into the chamber which peaked after 4 h. TNFα and IL-1β levels increased rapidly, reaching their maximum levels within 4 h. After 24 h, TNFα levels declined markedly and were undetectable at 48 and 96 h. TNFα mRNA levels in the sedimented cells followed a similar pattern. In contrast, IL-1β showed a more gradual decrease with levels significantly different from baseline still being present 96 h post-LPS challenge. Four consecutive daily injections of LPS into the chambers resulted in undetectable levels of TNFα in the chamber fluid, while significant levels of IL-1β were detected. These levels were significantly higher than the levels of IL-1β in the chamber fluid 96 h after a single injection and approximately 60% of the levels measured 24 h after a single intra-chamber injection of LPS. The results emphasize the difference between single and repeated exposure to LPS in vivo, and suggest a role for TNFα in the initial phase of the local inflammatory response and for IL-1β in the later phase.